Antimicrobial and Antiviral Nanofibers Halt Co‐Infection Spread via Nuclease‐Mimicry and Photocatalysis

Abstract The escalating spread of drug‐resistant bacteria and viruses is a grave concern for global health. Nucleic acids dominate the drug‐resistance and transmission of pathogenic microbes. Here, imidazolium‐type poly(ionic liquid)/porphyrin (PIL‐P) based electrospun nanofibrous membrane and its cerium (IV) ion complex (PIL‐P‐Ce) are developed. The obtained PIL‐P‐Ce membrane exhibits high and stable efficiency in eradicating various microorganisms (bacteria, fungi, and viruses) and decomposing microbial antibiotic resistance genes and viral nucleic acids under light. The nuclease‐mimetic and photocatalytic mechanisms of the PIL‐P‐Ce are elucidated. Co‐infection wound models in mice with methicillin‐resistant S. aureus and hepatitis B virus demonstrate that PIL‐P‐Ce integrate the triple effects of cationic polymer, photocatalysis, and nuclease‐mimetic activities. As revealed by proteomic analysis, PIL‐P‐Ce shows minimal phototoxicity to normal tissues. Hence, PIL‐P‐Ce has potential as a “green” wound dressing to curb the spread of drug‐resistant bacteria and viruses in clinical settings.


Figure S5 .
Figure S5.Physical property test of PIL-P-based membranes.A) Air permeability and water absorbability, B) water swelling rates (the inserted images show PIL-P-based membranes in PBS solution for 7 days), C) thermogravimetric analysis (TGA) curves, and D) protein absorbability and contact angles of PIL-P and PIL-P-Ce membranes.Data are mean ± s.d.N = 3.

Figure S6 .
Figure S6.In vitro biocompatibility of PIL-P-based membranes.A) Relative growth rate (RGR) and hemolysis rate of PIL-P and PIL-P-Ce in the absence or presence of 650 nm light (3.5 mW cm -2 ) for 30min.Hemocompatibility was assessed by hemolysis assays with fresh human red blood cells, and cytotoxicity by MTT assays using human fibroblasts (HDF) and human hepatocellular carcinoma cells (HepG2-NTCP).B) Images of HDF and HepG2-NTCP cells after 24 h incubation with the tested membranes post 30min light treatment.PET served as control.Scale bar, 100 μm.N = 3.

Figure S7 .
Figure S7.BNPP Decomposition by PIL-P-based membranes.A) Scheme of BNPP degradation by PIL-P-Ce under 650 nm light (3.5 mW cm -2 ).The insert photo displays 5 mM BNPP solutions after 4 h co-incubation with tested membranes, with or without 30 min light.The blank is BNPP solution without any membrane.B) Decomposition time course of 5 mM BNPP solutions by PIL-P-based membranes with or without 30 min lighting, quantified by absorbance of the p-nitrophenol product at 400 nm.A400 value is proportional to p-nitrophenol concentration.C) Initial velocity for PIL-P-Ce cleavage of different BNPP concentrations, with or without 30 min light.Insert table shows reaction kinetics parameters of the Michaelis-Menten equation for each sample.D) Recycling performance test for PIL-P-Ce decomposition of BNPP under 30 min light.Data are mean ± s.d.N = 3.

Figure S8 .
Figure S8.Impact of various factors for DNA degradation activity of PIL-P-Ce membranes.Electrophoretograms show bacterial plasmid cleavage efficiencies of PIL-P-Ce under different conditions in Figure 2G, with light for 30 min.

Figure S9 .
Figure S9.Intramolecular nucleophile activation mechanisms of DNA hydrolysis by Ce(IV) complex.Two computing modes for DNA hydrolysis, along with corresponding structures of reaction complex (RC) and product complex (PC) by DFT calculations are displayed in (A,B) and (C,D).Elements Ce, P, O, and H are colored with red, blue, azure, and white, respectively.Single-arrow lines indicate the reaction direction of participating groups, with red wavy lines highlighting the target P-O bond to be broken.Red dotted lines denote the length of P-O bond, blue dash lines signify distances of adjacent atoms, and other pertinent atomic distances are also marked (in Å).Bracketed values represent relative energies (in kcal/mol).

Figure S10 .
Figure S10.Antimicrobial activities of PIL-P-based membranes under light by colony assay.Three strains of MRSA, E. coli (Kan R ) and C. albicans were incubated with tested membranes under various exposure times to 650 nm light.PET was used as a control.

Figure S11 .
Figure S11.Antimicrobial activities of PIL-P-based membranes without light by colony assay.Three strains of MRSA, E. coli (Kan R ) and C. albicans were incubated with tested membranes at various exposure times without 650 nm light.PET was used as a control.

Figure S12 .
Figure S12.Transfer efficacies of ARGs in E. coli post-treatment with PIL-P-based membranes by colony assay.Plasmid from E. coli BL21 (Kan R ) was treated using PIL-P-based membranes, either with or without 30-min light exposure, and were subsequently mixed with E. coli DH5α (Amp R ) for transformation.Bacteria screened by dual antibiotics of kanamycin and ampicillin were counted to evaluate ARG transfer efficiencies, as presented in Figure 2H.PET was used as a control.

Figure S13 .
Figure S13.Anti-HBV activity of PIL-P-based membranes without light.A) Time course of HBV-DNA changes when viral sera contact control PET and PIL-P-based membranes without 650 nm light.B,C) HBcAg expression in HBV-infected hepatocytes for each group, detected via immunofluorescence assay without light.Scale bar, 50 μm.D-F) Levels of HBV-DNA and HBV antigens (HBsAg and HBeAg) in hepatocyte culture mediums for each group without light.PBS was used as a negative control.Data are mean ± s.d.N = 3. * P<0.05, **P<0.01,***P<0.001,PIL-P-based membranes are compared to PET control and analyzed by Student's t-test.

Figure S14 .
Figure S14.Images of wound treated with PIL-P-based membranes in a mouse model with bacterial-viral co-infection.Skin wounds were inoculated with a mixture of MRSA and HBV, and covered with tested PIL-P-based membranes for 4 h, with initial 30 min of 650 nm light exposure.Sterile gauze served as a control.At 4 h post application, tested membranes were immersed in sterile PBS for subsequent bacterial and viral assays.

Figure S15 .
Figure S15.Representative histological images of skin (Masson staining) and muscle (H&E staining) for PIL-P groups.Mouse back wounds were covered with PIL-P membranes and subjected to various light conditions: standard treatment (3.5 mW cm -2 for 30 min, PIL-P-L30),extended time (3.5 mW cm -2 for 240 min, PIL-P-L240), and enhanced power (70 mW cm -2 for 30 min, PIL-P-H30).Skin and muscle tissues from wound sites underwent for histological study.Sterile gauze treated at 3.5 mW cm -2 for 30 min was used as control.Scale bar, 100 μm.

Figure S16 .
Figure S16.Principal component analysis (PCA) and heatmap of proteomic data from skin and muscle samples.

Figure S17 .
Figure S17.PCA plot (A) and heatmap (B) of proteomic data across four skin groups.

Figure S18 .
Figure S18.PCA plot (A) and heatmap (B) of proteomic data across four muscle groups.

Figure S20 .
Figure S20.KEGG-enriched pathways and associated DEP fold changes for the skin comparison between control and P-L30, with the others depicted in Figure 6H.

Figure S21 .
Figure S21.Peak areas of major DEPs within KEGG pathways for the skin comparison between control and P-L30.Data are mean ± s.d.N = 3. ***P<0.001,analyzed by Student's ttest.